{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T17:48:09Z","timestamp":1790876889101,"version":"4.1.0"},"reference-count":54,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,5,22]],"date-time":"2020-05-22T00:00:00Z","timestamp":1590105600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"MINECO-FEDER, Madrid, Spain","award":["AGL2015-69735-R"],"award-info":[{"award-number":["AGL2015-69735-R"]}]},{"name":"Foundation Seneca, Murcia, Spain","award":["19892\/GERM\/15"],"award-info":[{"award-number":["19892\/GERM\/15"]}]},{"name":"MICIU\/FEDER, Madrid, Spain","award":["RTI2018-093525-B-I00"],"award-info":[{"award-number":["RTI2018-093525-B-I00"]}]},{"name":"The European Union's Horizon 2020 research and innovation program under the MSCA","award":["891663"],"award-info":[{"award-number":["891663"]}]},{"name":"Swedish Research Council FORMAS, Stockholm, Sweden","award":["Projects 2017-00946 and 2019-00288"],"award-info":[{"award-number":["Projects 2017-00946 and 2019-00288"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The infusion of boar seminal plasma (SP) before artificial insemination (AI) positively alters the expression of endometrial genes and pathways involved in embryo development. This study aimed to determine which transcriptome changes occur in preimplantation embryos in response to SP infusions during estrus. Postweaning estrus sows received 40-mL intrauterine infusions of either SP (N = 6) or BTS extender (control group; N = 6) 30 min before each of two post-cervical AIs. On Day 6, embryos were surgically collected and analyzed for differential gene expression. Microarray analysis of embryos revealed 210 annotated genes, differentially expressed (p-value &lt; 0.05 and fold change &lt;\/&gt; 2) in SP-blastocysts, compared to controls. Most of these genes were associated with biological, cellular, metabolic and developmental processes. The pathways enriched among the upregulated genes related to signal transduction, cellular processes and the endocrine system. Among altered genes involved in these pathways, the SP-group showed a conspicuous overexpression of ApoA-I, CDK1, MAPK1, SMAD2, PRKAA1 and RICTOR, with reported key roles in embryo development, implantation, or progression of pregnancy. In conclusion, the results demonstrate that SP infusions prior to AI upregulates the expression of embryo development related genes in Day 6 pig embryos.<\/jats:p>","DOI":"10.3390\/ijms21103662","type":"journal-article","created":{"date-parts":[[2020,5,22]],"date-time":"2020-05-22T10:18:18Z","timestamp":1590142698000},"page":"3662","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":31,"title":["Seminal Plasma Induces Overexpression of Genes Associated with Embryo Development and Implantation in Day-6 Porcine Blastocysts"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6811-0191","authenticated-orcid":false,"given":"Cristina A.","family":"Martinez","sequence":"first","affiliation":[{"name":"Department of Biomedical &amp; Clinical Sciences (BKV), BKH\/Obstetrics &amp; Gynaecology, Faculty of Medicine and Health Sciences, Link\u00f6ping University, SE-58185 Link\u00f6ping, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2010-5849","authenticated-orcid":false,"given":"Josep M.","family":"Cambra","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Campus de Ciencias de la Salud, Carretera Buenavista s\/n, 30120 El Palmar, Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6955-7750","authenticated-orcid":false,"given":"Maria A.","family":"Gil","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Campus de Ciencias de la Salud, Carretera Buenavista s\/n, 30120 El Palmar, Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5121-758X","authenticated-orcid":false,"given":"Inmaculada","family":"Parrilla","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Campus de Ciencias de la Salud, Carretera Buenavista s\/n, 30120 El Palmar, Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0120-354X","authenticated-orcid":false,"given":"Manuel","family":"Alvarez-Rodriguez","sequence":"additional","affiliation":[{"name":"Department of Biomedical &amp; Clinical Sciences (BKV), BKH\/Obstetrics &amp; Gynaecology, Faculty of Medicine and Health Sciences, Link\u00f6ping University, SE-58185 Link\u00f6ping, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5194-2124","authenticated-orcid":false,"given":"Heriberto","family":"Rodriguez-Martinez","sequence":"additional","affiliation":[{"name":"Department of Biomedical &amp; Clinical Sciences (BKV), BKH\/Obstetrics &amp; Gynaecology, Faculty of Medicine and Health Sciences, Link\u00f6ping University, SE-58185 Link\u00f6ping, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6202-5946","authenticated-orcid":false,"given":"Cristina","family":"Cuello","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Campus de Ciencias de la Salud, Carretera Buenavista s\/n, 30120 El Palmar, Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1260-9721","authenticated-orcid":false,"given":"Emilio A.","family":"Martinez","sequence":"additional","affiliation":[{"name":"Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), Campus de Ciencias de la Salud, Carretera Buenavista s\/n, 30120 El Palmar, Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,5,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.jri.2010.01.008","article-title":"Immune regulation of conception and embryo implantation-all about quality control?","volume":"85","author":"Robertson","year":"2010","journal-title":"J. Reprod. Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1007\/s00441-005-1127-3","article-title":"Seminal plasma and male factor signalling in the female reproductive tract","volume":"322","author":"Robertson","year":"2005","journal-title":"Cell Tissue Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1111\/j.1600-0897.2011.01033.x","article-title":"Seminal plasma proteins: What role do they play?","volume":"66","author":"Kvist","year":"2011","journal-title":"Am. J. Reprod. Immunol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1530\/jrf.0.1090029","article-title":"LH profile and advancement of ovulation after transcervical infusion of seminal plasma at different stages of oestrus in gilts","volume":"109","author":"Waberski","year":"1997","journal-title":"J. Reprod. Fertil."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1111\/j.1439-0531.1990.tb00682.x","article-title":"Advancement of Ovulation in the Sow Related to Seminal Plasma Application before Insemination","volume":"25","author":"Weitze","year":"1990","journal-title":"Reprod. Domest. Anim."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1530\/rep.1.01119","article-title":"Seminal plasma regulates ovarian progesterone production, leukocyte recruitment and follicular cell responses in the pig","volume":"132","author":"Jasper","year":"2006","journal-title":"Reproduction"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Waberski, D., Sch\u00e4fer, J., B\u00f6lling, A., Scheld, M., Henning, H., Hambruch, N., Schuberth, H.J., Pfarrer, C., Wrenzycki, C., and Hunter, R.H.F. (2018). Seminal plasma modulates the immune-cytokine network in the porcine uterine tissue and pre-ovulatory follicles. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0202654"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Alvarez-Rodriguez, M., Atikuzzaman, M., Venhoranta, H., Wright, D., and Rodriguez-Martinez, H. (2019). Expression of immune regulatory genes in the porcine internal genital tract is differentially triggered by spermatozoa and seminal plasma. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20030513"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1530\/jrf.0.0560129","article-title":"The role of the seminal vesicles, coagulating glands and prostate glands on the fertility and fecundity of mice","volume":"56","author":"Pang","year":"1979","journal-title":"J. Reprod. Fertil."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1111\/rda.12383","article-title":"Seminal fluid and immune adaptation for pregnancy\u2014Comparative biology in mammalian species","volume":"49","author":"Schjenken","year":"2014","journal-title":"Reprod. Domest. Anim."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1002\/mrd.20403","article-title":"Transmission of mitochondrial DNA in pigs and progeny derived from nuclear transfer of Meishan pig fibroblast cells","volume":"73","author":"Takeda","year":"2006","journal-title":"Mol. Reprod. Dev."},{"key":"ref_12","first-page":"399","article-title":"The use of different methods of oocyte activation for generation of porcine fibroblast cell nuclear-transferred embryos","volume":"10","author":"Samiec","year":"2010","journal-title":"Ann. Anim. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1530\/REP-15-0460","article-title":"BIX-01294 increases pig cloning efficiency by improving epigenetic reprogramming of somatic cell nuclei","volume":"151","author":"Huang","year":"2016","journal-title":"Reproduction"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"34980","DOI":"10.18632\/oncotarget.16507","article-title":"Dnmt1s in donor cells is a barrier to SCNT-mediated DNA methylation reprogramming in pigs","volume":"8","author":"Song","year":"2017","journal-title":"Oncotarget"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.theriogenology.2017.04.022","article-title":"In Vitro development and cytological quality of inter-species (porcine-bovine) cloned embryos are affected by trichostatin A-dependent epigenomic modulation of adult mesenchymal stem cells","volume":"97","author":"Opiela","year":"2017","journal-title":"Theriogenology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1002\/mrd.1120070107","article-title":"Reproduction in mice: The fate of spermatozoa not involved in fertilization","volume":"7","author":"Watson","year":"1983","journal-title":"Gamete Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1159\/000299080","article-title":"Influence of insemination on the implantation of transferred rat blastocysts","volume":"18","author":"Carp","year":"1984","journal-title":"Gynecol. Obstet. Investig."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1530\/rep.1.00160","article-title":"Seminal plasma regulates endometrial cytokine expression, leukocyte recruitment and embryo development in the pig","volume":"128","author":"Jasper","year":"2004","journal-title":"Reproduction"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fvets.2019.00465","article-title":"Seminal Plasma Modifies the Transcriptional Pattern of the Endometrium and Advances Embryo Development in Pigs","volume":"6","author":"Martinez","year":"2019","journal-title":"Front. Vet. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"99","DOI":"10.2527\/jas1975.40199x","article-title":"Freezing of boar spermatozoa: Fertilizing capacity with concentrated semen and a new thawing procedure","volume":"40","author":"Pursel","year":"1975","journal-title":"J. Anim. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/S0165-0378(01)00063-8","article-title":"A repertoire of cytokines in human seminal plasma","volume":"54","author":"Maegawa","year":"2002","journal-title":"J. Reprod. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1160","DOI":"10.1161\/ATVBAHA.118.312282","article-title":"Apolipoprotein A-I Protects Against Pregnancy-Induced Insulin Resistance in Rats","volume":"39","author":"Wu","year":"2019","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1055\/s-2007-1009242","article-title":"Serum levels of apolipoprotein A-I, A-II and HDL-cholesterol in second half of normal pregnancy and in pregnancy complicated by pre-eclampsia","volume":"21","author":"Rosing","year":"1989","journal-title":"Horm. Metab. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1139\/bcb-2015-0165","article-title":"Fructated apolipoprotein A-I exacerbates cellular senescence in human umbilical vein endothelial cells accompanied by impaired insulin secretion activity and embryo toxicity","volume":"94","author":"Park","year":"2016","journal-title":"Biochem. Cell Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.bbrc.2015.02.171","article-title":"Identification of stathmin 1 during peri-implantation period in mouse endometrium by a proteomics-based analysis","volume":"461","author":"Gou","year":"2015","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1016\/j.rbmo.2016.07.011","article-title":"Apolipoprotein A1 and heterogeneous nuclear ribonucleoprotein E1 implicated in the regulation of embryo implantation by inhibiting lipid peroxidation","volume":"33","author":"Jia","year":"2016","journal-title":"Reprod. Biomed. Online"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1016\/j.fertnstert.2011.05.049","article-title":"Identification of apolipoprotein A1 in the human embryonic secretome","volume":"96","author":"Mains","year":"2011","journal-title":"Fertil. Steril."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2476","DOI":"10.1093\/hmg\/dds061","article-title":"Cdk1, but not Cdk2, is the sole Cdk that is essential and sufficient to drive resumption of meiosis in mouse oocytes","volume":"21","author":"Adhikari","year":"2012","journal-title":"Hum. Mol. Genet."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1038\/nature06046","article-title":"Cdk1 is sufficient to drive the mammalian cell cycle","volume":"448","author":"Cerqueira","year":"2007","journal-title":"Nature"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1016\/j.devcel.2010.02.013","article-title":"Progressive Activation of CyclinB1-Cdk1 Coordinates Entry to Mitosis","volume":"18","author":"Gavet","year":"2010","journal-title":"Dev. Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1046\/j.1365-2443.2003.00680.x","article-title":"Essential role for ERK2 mitogen-activated protein kinase in placental development","volume":"8","author":"Hatano","year":"2003","journal-title":"Genes Cells"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"964","DOI":"10.1038\/sj.embor.embor939","article-title":"An essential function of the mitogen-activated protein kinase Erk2 in mouse trophoblast development","volume":"4","author":"Vella","year":"2003","journal-title":"EMBO Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1016\/j.mce.2013.08.024","article-title":"Epidermal growth factor stimulates proliferation and migration of porcine trophectoderm cells through protooncogenic protein kinase 1 and extracellular-signal-regulated kinases 1\/2 mitogen-activated protein kinase signal transduction cascades during early","volume":"381","author":"Jeong","year":"2013","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_34","first-page":"1","article-title":"Smad2 and Smad3 have differential sensitivity in relaying TGF\u03b2 signaling and inversely regulate early lineage specification","volume":"6","author":"Liu","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1038\/nri821","article-title":"CD4+CD25+ suppressor T cells: More questions than answers","volume":"2","author":"Shevach","year":"2002","journal-title":"Nat. Rev. Immunol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1038\/ni1037","article-title":"Regulatory T cells mediate maternal tolerance to the fetus","volume":"5","author":"Aluvihare","year":"2004","journal-title":"Nat. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.cmet.2008.11.008","article-title":"The Glycogen-Binding Domain on the AMPK \u03b2 Subunit Allows the Kinase to Act as a Glycogen Sensor","volume":"9","author":"McBride","year":"2009","journal-title":"Cell Metab."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.1038\/nature05828","article-title":"Energy-dependent regulation of cell structure by AMP-activated protein kinase","volume":"447","author":"Lee","year":"2007","journal-title":"Nature"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1152\/physrev.00026.2008","article-title":"LKB1 and AMPK family signaling: The intimate link between cell polarity and energy metabolism","volume":"89","author":"Jansen","year":"2009","journal-title":"Physiol. Rev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1016\/j.devcel.2006.08.013","article-title":"Multiallelic Disruption of the rictor Gene in Mice Reveals that mTOR Complex 2 Is Essential for Fetal Growth and Viability","volume":"11","author":"Shiota","year":"2006","journal-title":"Dev. Cell"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/srep17705","article-title":"Endothelial Rictor is crucial for midgestational development and sustained and extensive FGF2-induced neovascularization in the adult","volume":"5","author":"Aimi","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1158","DOI":"10.1242\/bio.20148730","article-title":"ATRX is required for maintenance of the neuroprogenitor cell pool in the embryonic mouse brain","volume":"3","author":"Ritchie","year":"2014","journal-title":"Biol. Open"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Garrick, D., Sharpe, J.A., Arkell, R., Dobbie, L., Smith, A.J.H., Wood, W.G., Higgs, D.R., and Gibbons, R.J. (2006). Loss of Atrx affects trophoblast development and the pattern of X-inactivation in extraembryonic tissues. PLoS Genet., 2.","DOI":"10.1371\/journal.pgen.0020058"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1245","DOI":"10.1126\/science.2884726","article-title":"Homeo boxes in the study of development","volume":"236","author":"Gehring","year":"1987","journal-title":"Science"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/0092-8674(92)90471-N","article-title":"Homeobox genes and axial patterning","volume":"68","author":"McGinnis","year":"1992","journal-title":"Cell"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3155","DOI":"10.1523\/JNEUROSCI.13-07-03155.1993","article-title":"Spatially restricted expression of Dlx-1, Dlx-2 (Tes-1), Gbx-2, and Wnt-3 in the embryonic day 12.5 mouse forebrain defines potential transverse and longitudinal segmental boundaries","volume":"13","author":"Bulfone","year":"1993","journal-title":"J. Neurosci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.theriogenology.2019.09.004","article-title":"Three-to-5-day weaning-to-estrus intervals do not affect neither efficiency of collection nor in vitro developmental ability of in vivo-derived pig zygotes","volume":"141","author":"Martinez","year":"2020","journal-title":"Theriogenology"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/srep10587","article-title":"Nonsurgical deep uterine transfer of vitrified, in vivo-derived, porcine embryos is as effective as the default surgical approach","volume":"5","author":"Martinez","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_49","unstructured":"Stringfellow, D.A., and Seidel, S.M. (1998). Photographic illustrations of embryo developmental stage and quality codes. Manual of the International Embryo Transfer Society, International Embryo Transfer Society (IETS)."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1093\/bioinformatics\/19.2.185","article-title":"A comparison of normalization methods for high density oligonucleotide array data based on variance and bias","volume":"19","author":"Bolstad","year":"2003","journal-title":"Bioinformatics"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1016\/j.tibtech.2005.05.011","article-title":"Pathways to the analysis of microarray data","volume":"23","author":"Curtis","year":"2005","journal-title":"Trends Biotechnol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1002\/0470857897.ch8","article-title":"The KEGG database","volume":"247","author":"Kanehisa","year":"2002","journal-title":"Novartis Found. Symp."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.1093\/bioinformatics\/btp101","article-title":"ClueGO: A Cytoscape plug-in to decipher functionally grouped gene ontology and pathway annotation networks","volume":"25","author":"Bindea","year":"2009","journal-title":"Bioinformatics"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1093\/nar\/29.9.e45","article-title":"A new mathematical model for relative quantification in real-time RT-PCR","volume":"29","author":"Pfaffl","year":"2001","journal-title":"Nucleic Acids Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/10\/3662\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:31:30Z","timestamp":1760175090000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/10\/3662"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5,22]]},"references-count":54,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2020,5]]}},"alternative-id":["ijms21103662"],"URL":"https:\/\/doi.org\/10.3390\/ijms21103662","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5,22]]}}}